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1
Recombinant Protein Expression, Crystallization, and Biophysical Studies of a Bacillus-conserved Nucleotide Pyrophosphorylase, BcMazG.芽孢杆菌保守核苷酸焦磷酸化酶BcMazG的重组蛋白表达、结晶及生物物理研究
J Vis Exp. 2017 May 16(123):55576. doi: 10.3791/55576.
2
Crystal structure of the Bacillus-conserved MazG protein, a nucleotide pyrophosphohydrolase.芽孢杆菌保守的核苷酸焦磷酸水解酶MazG蛋白的晶体结构
Biochem Biophys Res Commun. 2016 Mar 25;472(1):237-42. doi: 10.1016/j.bbrc.2016.02.097. Epub 2016 Feb 23.
3
Identification of a Bacillus anthracis specific indel in the yeaC gene and development of a rapid pyrosequencing assay for distinguishing B. anthracis from the B. cereus group.鉴定炭疽芽孢杆菌 yeaC 基因中的插入缺失并开发一种快速焦磷酸测序检测方法，用于区分炭疽芽孢杆菌和芽孢杆菌属。
J Microbiol Methods. 2011 Dec;87(3):278-85. doi: 10.1016/j.mimet.2011.08.015. Epub 2011 Sep 2.
4
What sets Bacillus anthracis apart from other Bacillus species?炭疽芽孢杆菌与其他芽孢杆菌属物种有何不同？
Annu Rev Microbiol. 2009;63:451-76. doi: 10.1146/annurev.micro.091208.073255.
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A spontaneous translational fusion of Bacillus cereus PlcR and PapR activates transcription of PlcR-dependent genes in Bacillus anthracis via binding with a specific palindromic sequence.蜡样芽孢杆菌PlcR和PapR的自发翻译融合通过与特定回文序列结合激活炭疽芽孢杆菌中PlcR依赖性基因的转录。
Infect Immun. 2004 Oct;72(10):5814-23. doi: 10.1128/IAI.72.10.5814-5823.2004.
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Complete sequence analysis of novel plasmids from emetic and periodontal Bacillus cereus isolates reveals a common evolutionary history among the B. cereus-group plasmids, including Bacillus anthracis pXO1.对来自产呕吐毒素蜡样芽孢杆菌和牙周炎蜡样芽孢杆菌分离株的新型质粒进行的全序列分析揭示了蜡样芽孢杆菌属质粒之间的共同进化史，包括炭疽芽孢杆菌pXO1。
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7
Genome sequence of Bacillus cereus and comparative analysis with Bacillus anthracis.蜡样芽孢杆菌的基因组序列及与炭疽芽孢杆菌的比较分析。
Nature. 2003 May 1;423(6935):87-91. doi: 10.1038/nature01582.
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The function of PlcR in Bacillus anthracis vaccine strain A16R.磷脂酶C调节蛋白（PlcR）在炭疽芽孢杆菌疫苗株A16R中的功能。
Yi Chuan. 2015 May;37(5):494-8. doi: 10.16288/j.yczz.15-040.
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Pathogenic Bacillus anthracis in the progressive gene losses and gains in adaptive evolution.致病性炭疽芽孢杆菌在适应性进化中的基因逐步丢失与获得。
BMC Bioinformatics. 2009 Jan 30;10 Suppl 1(Suppl 1):S3. doi: 10.1186/1471-2105-10-S1-S3.
10
Cereulide synthetase gene cluster from emetic Bacillus cereus: structure and location on a mega virulence plasmid related to Bacillus anthracis toxin plasmid pXO1.致吐性蜡样芽孢杆菌的cereulide合成酶基因簇：结构及其在与炭疽芽孢杆菌毒素质粒pXO1相关的一个巨型毒力质粒上的定位
BMC Microbiol. 2006 Mar 2;6:20. doi: 10.1186/1471-2180-6-20.

本文引用的文献

1
Structural and biochemical characterization of bacterial YpgQ protein reveals a metal-dependent nucleotide pyrophosphohydrolase.细菌YpgQ蛋白的结构与生化特性揭示了一种金属依赖性核苷酸焦磷酸水解酶。
J Struct Biol. 2016 Jul;195(1):113-22. doi: 10.1016/j.jsb.2016.04.002. Epub 2016 Apr 7.
2
Crystal structure of the Bacillus-conserved MazG protein, a nucleotide pyrophosphohydrolase.芽孢杆菌保守的核苷酸焦磷酸水解酶MazG蛋白的晶体结构
Biochem Biophys Res Commun. 2016 Mar 25;472(1):237-42. doi: 10.1016/j.bbrc.2016.02.097. Epub 2016 Feb 23.
3
LmbE proteins from Bacillus cereus are de-N-acetylases with broad substrate specificity and are highly similar to proteins in Bacillus anthracis.蜡样芽胞杆菌 LmbE 蛋白是一种具有广泛底物特异性的去乙酰化酶，与炭疽芽胞杆菌中的蛋白质高度相似。
FEBS J. 2010 Jul;277(13):2740-53. doi: 10.1111/j.1742-4658.2010.07691.x. Epub 2010 May 19.
4
Crystal structure of Escherichia coli MazG, the regulator of nutritional stress response.大肠杆菌MazG（营养应激反应调节因子）的晶体结构
J Biol Chem. 2008 May 30;283(22):15232-40. doi: 10.1074/jbc.M800479200. Epub 2008 Mar 18.
5
A putative house-cleaning enzyme encoded within an integron array: 1.8 A crystal structure defines a new MazG subtype.整合子阵列中编码的一种假定的 house-cleaning 酶：1.8 Å 晶体结构定义了一种新的 MazG 亚型。
Mol Microbiol. 2007 Nov;66(3):610-21. doi: 10.1111/j.1365-2958.2007.05932.x. Epub 2007 Sep 24.
6
MazG -- a regulator of programmed cell death in Escherichia coli.MazG——大肠杆菌中程序性细胞死亡的调节因子。
Mol Microbiol. 2006 Jan;59(2):590-601. doi: 10.1111/j.1365-2958.2005.04956.x.
7
House cleaning, a part of good housekeeping.房屋清洁是良好家政管理的一部分。
Mol Microbiol. 2006 Jan;59(1):5-19. doi: 10.1111/j.1365-2958.2005.04950.x.
8
Protein production and crystallization at the joint center for structural genomics.结构基因组学联合中心的蛋白质生产与结晶
J Struct Funct Genomics. 2005;6(2-3):71-9. doi: 10.1007/s10969-005-2897-2.
9
Dimeric dUTPases, HisE, and MazG belong to a new superfamily of all-alpha NTP pyrophosphohydrolases with potential "house-cleaning" functions.二聚体脱氧尿苷三磷酸酶、组氨酸酶E和MazG属于一个全新的全α-型NTP焦磷酸水解酶超家族，具有潜在的“清理”功能。
J Mol Biol. 2005 Mar 25;347(2):243-55. doi: 10.1016/j.jmb.2005.01.030. Epub 2005 Jan 27.
10
The crystal structure of a complex of Campylobacter jejuni dUTPase with substrate analogue sheds light on the mechanism and suggests the "basic module" for dimeric d(C/U)TPases.空肠弯曲菌dUTPase与底物类似物复合物的晶体结构揭示了其作用机制，并为二聚体d（C/U）TPases提出了“基本模块”。
J Mol Biol. 2004 Oct 1;342(5):1583-97. doi: 10.1016/j.jmb.2004.07.050.

芽孢杆菌保守核苷酸焦磷酸化酶BcMazG的重组蛋白表达、结晶及生物物理研究

Recombinant Protein Expression, Crystallization, and Biophysical Studies of a Bacillus-conserved Nucleotide Pyrophosphorylase, BcMazG.

作者信息

Kim Meong Il, Lee Choongdeok, Hong Minsun

机构信息

Division of Biological Science and Technology, Yonsei University.

Division of Biological Science and Technology, Yonsei University;

出版信息

J Vis Exp. 2017 May 16(123):55576. doi: 10.3791/55576.

DOI:10.3791/55576
PMID:28570540
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5607965/
Abstract

To overcome safety restrictions and regulations when studying genes and proteins from true pathogens, their homologues can be studied. Bacillus anthracis is an obligate pathogen that causes fatal inhalational anthrax. Bacillus cereus is considered a useful model for studying B. anthracis due to its close evolutionary relationship. The gene cluster ba1554 - ba1558 of B. anthracis is highly conserved with the bc1531- bc1535 cluster in B. cereus, as well as with the bt1364-bt1368 cluster in Bacillus thuringiensis, indicating the critical role of the associated genes in the Bacillus genus. This manuscript describes methods to prepare and characterize a protein product of the first gene (ba1554) from the gene cluster in B. anthracis using a recombinant protein of its ortholog in B. cereus, bc1531.

摘要

为了在研究来自真正病原体的基因和蛋白质时克服安全限制和规定，可以研究它们的同源物。炭疽芽孢杆菌是一种导致致命吸入性炭疽的专性病原体。蜡样芽孢杆菌由于其密切的进化关系，被认为是研究炭疽芽孢杆菌的有用模型。炭疽芽孢杆菌的基因簇ba1554 - ba1558与蜡样芽孢杆菌的bc1531 - bc1535簇以及苏云金芽孢杆菌的bt1364 - bt1368簇高度保守，这表明相关基因在芽孢杆菌属中具有关键作用。本手稿描述了使用蜡样芽孢杆菌中其直系同源基因bc1531的重组蛋白来制备和表征炭疽芽孢杆菌基因簇中第一个基因（ba1554）的蛋白质产物的方法。